Yang Wenhua, Gu Qiuya, Zhou Jianli, Liu Xiaobo, Yu Xiaobin
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214000, China.
Guizhou Province Key Laboratory of Fermentation Engineering and Biopharmacy, School of Liquor and Food Engineering, Guizhou University, Guiyang 550003, China.
Foods. 2023 Jan 20;12(3):496. doi: 10.3390/foods12030496.
Deep eutectic solvents (DES), as a green alternative to traditional organic solvents in biocatalysis, not only activate proteins but even increase the efficiency of enzymatic reactions. Here, DES were used in a combinatorial enzyme-catalyzed system containing β-glucosidase BGLAt and β-galactosidase BGALAo to produce deglycosylated ginsenosides (De-g) from ginseng extracts (GE). The results showed that DES prepared with betaine and ethylene glycol (molar ratio, 1:2) could significantly stimulate the activity of the combinatorial enzymes as well as improve the acid resistance and temperature stability. The DES-based combinatorial enzyme-catalyzed system could convert 5 g of GE into 1.24 g of De-g (F, F, 20 ()-PPT, and CK) at 24 h, which was 1.1 times that of the buffer sample. As confirmed by the spectral data, the changes in the conformations of the combinatorial enzymes were more favorable for the binding reaction with the substrates. Moreover, the constructed DES-based aqueous two-phase system enabled the recovery of substantial amounts of DES and De-g from the top phase. These results demonstrated that DES shows great application as a reaction solvent for the scale-up production of De-g and provide insights for the green extraction of natural products.
深共熔溶剂(DES)作为生物催化中传统有机溶剂的绿色替代品,不仅能激活蛋白质,甚至还能提高酶促反应的效率。在此,DES被用于包含β-葡萄糖苷酶BGLAt和β-半乳糖苷酶BGALAo的组合酶催化体系中,以从人参提取物(GE)中生产去糖基化人参皂苷(De-g)。结果表明,由甜菜碱和乙二醇(摩尔比为1:2)制备的DES能显著刺激组合酶的活性,同时提高其耐酸性和热稳定性。基于DES的组合酶催化体系在24小时内可将5克GE转化为1.24克De-g(F、F、20()-PPT和CK),是缓冲液样品的1.1倍。光谱数据证实,组合酶构象的变化更有利于与底物的结合反应。此外,构建的基于DES的双水相体系能够从上层相中回收大量的DES和De-g。这些结果表明,DES作为De-g规模化生产的反应溶剂具有巨大的应用潜力,并为天然产物的绿色提取提供了思路。